Myelodysplastic syndromes (MDS) are myeloid malignancies with heterogeneous genotypes and phenotypes, characterized by ineffective haematopoiesis and a high risk of progression towards acute myeloid leukaemia (AML). Prognosis for patients treated with hypomethylating agents (HMAs), as is azacytidine, the main drug used as frontline therapy for MDS is mostly based on cytogenetics and next generation sequencing (NGS) of the initial myeloid clone. Although the critical influence of the epigenetic landscape upon cancer cells survival and development as well on tumour environment establishment is currently recognized and approached within current clinical practice in MDS, the heterogenous response of the patients to epigenetic therapy is suggesting a more complex mechanism of action, as is the case of RNA methylation. In this sense, the newly emerging field of epitranscriptomics could provide a more comprehensive perspective upon the modulation of gene expression in malignancies, as is the proof-of-concept of MDS. We initially did RNA methylation sequencing on MDS patients (n = 6) treated with azacytidine and compared responders with non-responders. Afterwards, the genes identified were assessed in vitro and afterwards validated on a larger cohort of MDS patients treated with azacytidine (n = 58). Our data show that a more accurate prognosis could be based on analysing the methylome and thus we used methylation sequencing to differentially split high-grade MDS patients with identical demographical and cytogenetic features, between azacytidine responders and non-responders.

• MeSH
• antimetabolity antitumorózní terapeutické užití   farmakologie   MeSH
• azacytidin * farmakologie   terapeutické užití   MeSH
• epigeneze genetická účinky léků   MeSH
• lidé středního věku MeSH
• lidé MeSH
• methylace RNA MeSH
• metylace DNA * účinky léků   MeSH
• myelodysplastické syndromy * genetika   farmakoterapie   patologie   MeSH
• prognóza MeSH
• sekvenční analýza RNA MeSH
• senioři nad 80 let MeSH
• senioři MeSH
• stanovení celkové genové exprese MeSH
• transkriptom genetika   účinky léků   MeSH
• vysoce účinné nukleotidové sekvenování MeSH
• Check Tag
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• senioři nad 80 let MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH

Current standard-of-care systemic therapy options for locally advanced and metastatic bladder cancer (BC), which are predominantly based on cisplatin-gemcitabine combinations, are limited by significant treatment failure rates and frailty-based patient ineligibility. We previously addressed the urgent clinical need for better-tolerated BC therapeutic strategies using a drug screening approach, which identified outstanding antineoplastic activity of clofarabine in preclinical models of BC. To further assess clofarabine as a potential BC therapy component, we conducted head-to-head comparisons of responses to clofarabine versus gemcitabine in preclinical in vitro and in vivo models of BC, complemented by in silico analyses. In vitro data suggest a distinct correlation between the two antimetabolites, with higher cytotoxicity of gemcitabine, especially against several nonmalignant cell types, including keratinocytes and endothelial cells. Accordingly, tolerance of clofarabine (oral or intraperitoneal application) was distinctly better than for gemcitabine (intraperitoneal) in patient-derived xenograft models of BC. Clofarabine also exhibited distinctly superior anticancer efficacy, even at dosing regimens optimized for gemcitabine. Neither complete remission nor cure, both of which were observed with clofarabine, were achieved with any tolerable gemcitabine regimen. Taken together, our findings demonstrate that clofarabine has a better therapeutic window than gemcitabine, further emphasizing its potential as a candidate for drug repurposing in BC. PATIENT SUMMARY: We compared the anticancer activity of clofarabine, a drug used for treatment of leukemia but not bladder cancer, and gemcitabine, a drug currently used for chemotherapy against bladder cancer. Using cell cultures and mouse models, we found that clofarabine was better tolerated and more efficacious than gemcitabine, and even cured implanted tumors in mouse models. Our results suggest that clofarabine, alone or in combination schemes, might be superior to gemcitabine for the treatment of bladder cancer.

• MeSH
• antimetabolity antitumorózní terapeutické užití   farmakologie   MeSH
• deoxycytidin * analogy a deriváty   farmakologie   terapeutické užití   MeSH
• gemcitabin * MeSH
• klofarabin * terapeutické užití   farmakologie   MeSH
• lidé MeSH
• myši MeSH
• nádorové buněčné linie MeSH
• nádory močového měchýře * farmakoterapie   patologie   MeSH
• xenogenní modely - testy antitumorózní aktivity MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• myši MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• srovnávací studie MeSH

• Klíčová slova
• venetoklax,
• MeSH
• analýza přežití MeSH
• antimetabolity antitumorózní farmakologie   terapeutické užití   MeSH
• antitumorózní látky farmakologie   terapeutické užití   MeSH
• azacytidin farmakologie   terapeutické užití   MeSH
• bicyklické sloučeniny heterocyklické farmakologie   terapeutické užití   MeSH
• decitabin farmakologie   terapeutické užití   MeSH
• klinická studie jako téma MeSH
• lidé MeSH
• myelodysplastické syndromy * farmakoterapie   MeSH
• protokoly antitumorózní kombinované chemoterapie MeSH
• sulfonamidy farmakologie   terapeutické užití   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• práce podpořená grantem MeSH
• přehledy MeSH

3-dimensional (3D) cell cultures are being increasingly recognized as physiologically more relevant in vitro models than traditional monolayer cultures, because they better mimic in vivo-like microenvironment, cell-cell and cell-extracellular matrix interactions. Nevertheless, the broader use of 3D models might be limited by requirements for special consumables, equipment, or skills for 3D cell cultures, and by their limited throughput and scalability. In this study, we optimized and adapted a commercially available agarose-micromolding technique to produce scaffold-free spheroid cultures. Brightfield microscopy was used for routine nondestructive and noninvasive evaluation of spheroid formation and growth. The workflow is compatible with manual, as well as high speed automated microscopic image acquisition, and it is supplemented with an in-house developed macro 'Spheroid_Finder' for open source software Fiji to facilitate rapid automated image analysis. This protocol was used to characterize and quantify spheroid formation and growth of two different hepatic cell lines, hTERT immortalized, but non-cancerous, adult human liver stem cell line HL1-hT1, and human hepatocellular carcinoma cell line HepG2, as well as their responses to a model antiproliferative and cytotoxic agent, 5-fluorouracil. The complete protocol provides a simple and ready-to-use solution to initiate scaffold-free spheroid cultures in any laboratory with standard equipment for mammalian in vitro cell culture work. Thus, it allows to increase throughput and scale of spheroid culture experiments, which can be greatly utilized in different areas of biomedical, pharmaceutical and toxicological research.

• MeSH
• antimetabolity antitumorózní farmakologie   toxicita   MeSH
• buněčné kultury MeSH
• buněčné sféroidy MeSH
• buňky Hep G2 MeSH
• časové faktory MeSH
• fluorouracil farmakologie   toxicita   MeSH
• hepatocelulární karcinom farmakoterapie   metabolismus   patologie   MeSH
• játra účinky léků   metabolismus   patologie   MeSH
• kmenové buňky účinky léků   metabolismus   patologie   MeSH
• léky antitumorózní - screeningové testy MeSH
• lidé MeSH
• nádory jater farmakoterapie   metabolismus   patologie   MeSH
• proliferace buněk účinky léků   MeSH
• průběh práce MeSH
• rychlé screeningové testy * MeSH
• testy toxicity MeSH
• viabilita buněk účinky léků   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH

The natural behavior of mesenchymal stem cells (MSCs) and their exosomes in targeting tumors is a promising approach for curative therapy. Human tumor tropic mesenchymal stem cells (MSCs) isolated from various tissues and MSCs engineered to express the yeast cytosine deaminase::uracil phosphoribosyl transferase suicide fusion gene (yCD::UPRT-MSCs) released exosomes in conditional medium (CM). Exosomes from all tissue specific yCD::UPRT-MSCs contained mRNA of the suicide gene in the exosome's cargo. When the CM was applied to tumor cells, the exosomes were internalized by recipient tumor cells and in the presence of the prodrug 5-fluorocytosine (5-FC) effectively triggered dose-dependent tumor cell death by endocytosed exosomes via an intracellular conversion of the prodrug 5-FC to 5-fluorouracil. Exosomes were found to be responsible for the tumor inhibitory activity. The presence of microRNAs in exosomes produced from naive MSCs and from suicide gene transduced MSCs did not differ significantly. MicroRNAs from yCD::UPRT-MSCs were not associated with therapeutic effect. MSC suicide gene exosomes represent a new class of tumor cell targeting drug acting intracellular with curative potential.

• MeSH
• antimetabolity antitumorózní metabolismus   farmakologie   MeSH
• cytosindeaminasa genetika   metabolismus   MeSH
• exozómy genetika   metabolismus   MeSH
• flucytosin metabolismus   MeSH
• fluorouracil metabolismus   farmakologie   MeSH
• fungální proteiny genetika   metabolismus   MeSH
• genetická terapie metody   MeSH
• kvasinky genetika   metabolismus   MeSH
• lidé MeSH
• mezenchymální kmenové buňky metabolismus   MeSH
• nádorové buněčné linie MeSH
• nádory prsu genetika   metabolismus   patologie   MeSH
• pentosyltransferasy genetika   metabolismus   MeSH
• prekurzory léčiv metabolismus   MeSH
• proliferace buněk účinky léků   genetika   MeSH
• transgeny sebevražedné genetika   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

The first-line chemotherapy of colorectal cancer (CRC), besides surgery, comprises administration of 5-Fluorouracil (5FU). Apart from cytotoxic effect on cancer cells, 5FU may also cause adverse side effects. Ganoderma Lucidum (GLC) is a mushroom used in Traditional Eastern Medicine. We propose that natural compounds, particularly GLC extracts, may sensitize cancer cells to conventional chemotherapeutics. This combination therapy could lead to more selective cancer cell death and may improve the response to the therapy and diminish the adverse effects of anticancer drugs. Here we demonstrate that GLC induced oxidative DNA damage selectively in colorectal cancer cell lines, whereas it protected non-malignant cells from the accumulation of reactive oxygen species. Accumulation of DNA damage caused sensitization of cancer cells to 5FU resulting in improved anticancer effect of 5FU. The results obtained in colorectal cell lines were confirmed in in vivo study: GLC co-treatment with 5FU increased the survival of treated mice and reduced the tumor volume in comparison with group treated with 5FU alone. Combination of conventional chemotherapeutics and natural compounds is a promising approach, which may reduce the effective curative dose of anticancer drugs, suppress their adverse effects and ultimately lead to better quality of life of CRC patients.

• MeSH
• adenokarcinom farmakoterapie   patologie   MeSH
• antimetabolity antitumorózní farmakologie   terapeutické užití   MeSH
• buněčné dělení účinky léků   MeSH
• DNA nádorová účinky léků   MeSH
• fluorouracil farmakologie   terapeutické užití   MeSH
• invazivní růst nádoru MeSH
• kolorektální nádory farmakoterapie   patologie   MeSH
• kometový test MeSH
• léky antitumorózní - screeningové testy MeSH
• myši inbrední BALB C MeSH
• myši MeSH
• nádorové buněčné linie MeSH
• oxidační stres MeSH
• poškození DNA * MeSH
• reaktivní formy kyslíku metabolismus   MeSH
• reishi chemie   MeSH
• rostlinné extrakty izolace a purifikace   farmakologie   MeSH
• synergismus léků MeSH
• testy nádorových kmenových buněk MeSH
• tumor burden účinky léků   MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Introduction of small-molecule inhibitors of B-cell receptor signaling and BCL2 protein significantly improves therapeutic options in chronic lymphocytic leukemia. However, some patients suffer from adverse effects mandating treatment discontinuation, and cases with TP53 defects more frequently experience early progression of the disease. Development of alternative therapeutic approaches is, therefore, of critical importance. Here we report details of the anti-chronic lymphocytic leukemia single-agent activity of MU380, our recently identified potent, selective, and metabolically robust inhibitor of checkpoint kinase 1. We also describe a newly developed enantioselective synthesis of MU380, which allows preparation of gram quantities of the substance. Checkpoint kinase 1 is a master regulator of replication operating primarily in intra-S and G2/M cell cycle checkpoints. Initially tested in leukemia and lymphoma cell lines, MU380 significantly potentiated efficacy of gemcitabine, a clinically used inducer of replication stress. Moreover, MU380 manifested substantial single-agent activity in both TP53-wild type and TP53-mutated leukemia and lymphoma cell lines. In chronic lymphocytic leukemia-derived cell lines MEC-1, MEC-2 (both TP53-mut), and OSU-CLL (TP53-wt) the inhibitor impaired cell cycle progression and induced apoptosis. In primary clinical samples, MU380 used as a single-agent noticeably reduced the viability of unstimulated chronic lymphocytic leukemia cells as well as those induced to proliferate by anti-CD40/IL-4 stimuli. In both cases, effects were comparable in samples harboring p53 pathway dysfunction (TP53 mutations or ATM mutations) and TP53-wt/ATM-wt cells. Lastly, MU380 also exhibited significant in vivo activity in a xenotransplant mouse model (immunodeficient strain NOD-scid IL2Rγnull ) where it efficiently suppressed growth of subcutaneous tumors generated from MEC-1 cells.

• MeSH
• antimetabolity antitumorózní farmakologie   MeSH
• apoptóza MeSH
• buněčný cyklus MeSH
• checkpoint kinasa 1 antagonisté a inhibitory   MeSH
• chemorezistence účinky léků   MeSH
• chronická lymfatická leukemie farmakoterapie   genetika   patologie   MeSH
• deoxycytidin analogy a deriváty   farmakologie   MeSH
• inhibitory proteinkinas farmakologie   MeSH
• lidé MeSH
• mutace * MeSH
• myši inbrední NOD MeSH
• myši SCID MeSH
• myši MeSH
• nádorové biomarkery genetika   MeSH
• nádorové buňky kultivované MeSH
• nádorový supresorový protein p53 genetika   MeSH
• piperidiny farmakologie   MeSH
• proliferace buněk MeSH
• pyrazoly farmakologie   MeSH
• pyrimidiny farmakologie   MeSH
• regulace genové exprese u nádorů účinky léků   MeSH
• synergismus léků * MeSH
• xenogenní modely - testy antitumorózní aktivity MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• myši MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

• MeSH
• analýza přežití MeSH
• antimetabolity antitumorózní farmakologie   terapeutické užití   MeSH
• antracykliny farmakologie   terapeutické užití   MeSH
• cisplatina farmakologie   terapeutické užití   MeSH
• docetaxel farmakologie   terapeutické užití   MeSH
• inhibitory angiogeneze farmakologie   terapeutické užití   MeSH
• irinotekan farmakologie   terapeutické užití   MeSH
• lidé MeSH
• metastázy nádorů farmakoterapie   MeSH
• nádory žaludku * farmakoterapie   patologie   MeSH
• nežádoucí účinky léčiv MeSH
• oxaliplatin farmakologie   terapeutické užití   MeSH
• paliativní péče metody   MeSH
• protokoly antitumorózní kombinované chemoterapie normy   MeSH
• stupeň nádoru MeSH
• trastuzumab farmakologie   terapeutické užití   MeSH
• vaskulární endoteliální růstový faktor A antagonisté a inhibitory   MeSH
• Check Tag
• lidé MeSH

BACKGROUND: Azacitidine (AZA) is a nucleoside analog used for treatment of myelodysplasia and the prediction of AZA responsiveness is important for the therapy management. METHODS: Using microarrays and reverse-transcription quantitative-PCR, we analyzed microRNA (miRNA) expression in bone marrow CD34+ cells of 27 patients with higher-risk myelodysplastic syndromes or acute myeloid leukemia with myelodysplasia-related changes before and during AZA treatment. RESULTS: At baseline, we found that future overall response rate was significantly higher in patients with upregulated miR-17-3p and downregulated miR-100-5p and miR-133b. Importantly, the high level of miR-100-5p at baseline was associated with shorter overall survival (HR = 4.066, P= 0.008). After AZA treatment, we observed deregulation of 30 miRNAs in responders (including downregulation of miR-10b-5p, miR-15a-5p/b-5p, miR-24-3p, and miR-148b-3p), while their levels remained unchanged in non-responders. CONCLUSIONS: Our study demonstrates that responders and non-responders have distinct miRNA patterns and that the level of specific miRNAs before therapy may predict the efficacy of AZA treatment.

• MeSH
• akutní myeloidní leukemie farmakoterapie   genetika   patologie   MeSH
• antimetabolity antitumorózní farmakologie   terapeutické užití   MeSH
• azacytidin farmakologie   terapeutické užití   MeSH
• lidé MeSH
• mikro RNA metabolismus   MeSH
• myelodysplastické syndromy farmakoterapie   genetika   patologie   MeSH
• senioři MeSH
• Check Tag
• lidé MeSH
• mužské pohlaví MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH

BACKGROUND: Activating KRAS mutations are reported in up to 90% of pancreatic cancers. Refametinib potently inhibits MEK1/2, part of the MAPK signaling pathway. This phase I/II study evaluated the safety and efficacy of refametinib plus gemcitabine in patients with advanced pancreatic cancer. METHODS: Phase I comprised dose escalation, followed by phase II expansion. Refametinib and gemcitabine plasma levels were analyzed for pharmacokinetics. KRAS mutational status was determined from circulating tumor DNA. RESULTS: Ninety patients overall received treatment. The maximum tolerated dose was refametinib 50 mg twice daily plus standard gemcitabine (1000 mg/m2 weekly). The combination was well tolerated, with no pharmacokinetic interaction. Treatment-emergent toxicities included thrombocytopenia, fatigue, anemia, and edema. The objective response rate was 23% and the disease control rate was 73%. Overall response rate, disease control rate, progression-free survival, and overall survival were higher in patients without detectable KRAS mutations (48% vs. 28%, 81% vs. 69%, 8.8 vs. 5.3 months, and 18.2 vs. 6.6 months, respectively). CONCLUSION: Refametinib plus gemcitabine was well tolerated, with a promising objective response rate, and had an acceptable safety profile and no pharmacokinetic interaction. There was a trend towards improved outcomes in patients without detectable KRAS mutations that deserves future investigation.

• MeSH
• antimetabolity antitumorózní farmakokinetika   farmakologie   terapeutické užití   MeSH
• deoxycytidin analogy a deriváty   farmakokinetika   farmakologie   terapeutické užití   MeSH
• difenylamin analogy a deriváty   farmakokinetika   farmakologie   terapeutické užití   MeSH
• lidé středního věku MeSH
• lidé MeSH
• nádory slinivky břišní farmakoterapie   MeSH
• přežití po terapii bez příznaků nemoci MeSH
• sulfonamidy farmakokinetika   farmakologie   terapeutické užití   MeSH
• výsledek terapie MeSH
• Check Tag
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• klinické zkoušky, fáze I MeSH
• klinické zkoušky, fáze II MeSH
• multicentrická studie MeSH
• práce podpořená grantem MeSH